Structural investigations by NMR or X-ray crystallography require a homogenous RNA preparation. However, in vitro transcription-based RNA synthesis with T7 RNA polymerase produces heterogenous RNA at both 5' and 3' ends especially if prepared under the commonly used class III G-initiating phi6.5 promotor.
The combination of
| Product | Cat.-No. | Amount | Price | Application | Average yield / react.* |
|---|---|---|---|---|---|
| HighYield T7 P&L RNA Synthesis Kit | RNT-201 | 50 reactions x 20 µl | 195,00 € | Optimized kit for high yield RNA synthesis | 140 - 160 µg |
| T7 P&L RNA Polymerase HC | RNT-018 | 50 µl (2 µg/µl) | 365,00 € | Single enzyme for individual assay set-ups | n/a |
| HighYield T7 P&L RNA NMR Kit (5F-UTP) | RNT-202-S | 15 reactions x 20 µl | 395,00 € | Optimized kit for preparation of 5-Fluoro-modified RNA | 80 µg |
| RNT-202-L | 50 reactions x 20 µl | 895,00 € | |||
| 5-Fluoro-UTP (5F-UTP) | NU-1115S | 10 µl (100 mM) | 157,60 € | Single nucleotide for individual assay set-up | n/a |
| NU-1115L | 5 x 10 µl (100 mM) | 461,70 € | |||
| HighYield T7 P&L RNA NMR Kit (2F-ATP) | RNT-203-S | 15 reactions x 20 µl | 535,00 € | Optimized kit for preparation of 2-Fluoro-modified RNA | 80 µg |
| RNT-203-L | 50 reactions x 20 µl | 1.140,00 € | |||
| 2-Fluoro-ATP (2F-ATP) | NU-145S | 10 µl (100 mM) | 208,50 € | Single nucleotide for individual assay set-up | n/a |
| NU-145L | 5 x 10 µl (100 mM) | 604,60 € |
[1] Coleman et al. (2004) Superior 5‘ homogeneity of RNA from ATP-initiated transcription under T7 phi 2.5 promoter. Nucleic Acids Research 32 (1):e14.
[2] Guillerez et al. (2005) A mutation in T7 RNA polymerase that facilitates promoter clearance. Natl. Acad. Sci. U.S.A. 102:5958.
[3] Salvail-Lacoste et al. (2018) Affinity purification of T7 RNA transcripts with homogeneous ends using ARiBo and CRISPR tags. RNA 19:1003.
